U.S. patent number 10,492,886 [Application Number 15/313,378] was granted by the patent office on 2019-12-03 for ink for marking a tissue specimen.
This patent grant is currently assigned to VECTOR SURGICAL, LLC. The grantee listed for this patent is Janet L. F. Phillips, Vector Surgical, LLC. Invention is credited to Scott E. Moore, Janet L. F. Phillips.
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United States Patent |
10,492,886 |
Phillips , et al. |
December 3, 2019 |
Ink for marking a tissue specimen
Abstract
An ink composition for marking a tissue specimen is provided.
The ink composition has a general formula of 30.0 wt. % to 54.0 wt.
% of an alkali soluble styrene; 0.3 wt. % to 1.7 wt. % ethyl
hydroxyethyl cellulose; 8.0 wt. % to 35.0 wt. % colorant; 0.0 wt. %
to 13.0 wt. % pigment; 23.0 wt. % to 47.0 wt. % deionized water;
0.35 wt. % to 1.65% defoamer and 0.1 wt. % to 1.1 wt. %
preservative. The inks in accordance with the invention may have a
peak transmission in the visible spectrum at a wavelength of from
322 nm to 716 nm or having a lineal UV-Vis spectrum with no visible
peak transmission between 250 nm and 950 nm. When applied to a
tissue specimen the ink compositions do not bleed onto adjacent
tissue margins. When view under a microscope the color of the ink
compositions can be distinguished from each other.
Inventors: |
Phillips; Janet L. F.
(Oconomowoc, WI), Moore; Scott E. (Howell, MI) |
Applicant: |
Name |
City |
State |
Country |
Type |
Phillips; Janet L. F.
Vector Surgical, LLC |
Oconomowoc
Waukesha |
WI
WI |
US
US |
|
|
Assignee: |
VECTOR SURGICAL, LLC (Waukesha,
WI)
|
Family
ID: |
57546314 |
Appl.
No.: |
15/313,378 |
Filed: |
June 17, 2016 |
PCT
Filed: |
June 17, 2016 |
PCT No.: |
PCT/US2016/038088 |
371(c)(1),(2),(4) Date: |
November 22, 2016 |
PCT
Pub. No.: |
WO2016/205657 |
PCT
Pub. Date: |
December 22, 2016 |
Prior Publication Data
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|
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Document
Identifier |
Publication Date |
|
US 20170189135 A1 |
Jul 6, 2017 |
|
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
|
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62182153 |
Jun 19, 2015 |
|
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
G01N
1/30 (20130101); A61B 90/92 (20160201); C09D
11/00 (20130101); C09D 11/54 (20130101); A61B
90/90 (20160201); G01N 1/28 (20130101) |
Current International
Class: |
C09D
11/00 (20140101); C09D 11/54 (20140101); A61B
90/90 (20160101); A61B 90/92 (20160101); G01N
1/28 (20060101); G01N 1/30 (20060101) |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
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101306943 |
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Nov 2008 |
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CN |
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103278649 |
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Sep 2013 |
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CN |
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103305053 |
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Sep 2013 |
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CN |
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104335362 |
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Feb 2015 |
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CN |
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2388567 |
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Nov 2011 |
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EP |
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5035496 |
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Sep 2012 |
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JP |
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WO-2014062227 |
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Apr 2014 |
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WO |
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Other References
"Conversion between Stormer Viscometer Krebs Units and Viscosity
Cup Drain Time"; Paul N. Gardner Company, Inc.; Jan. 1999;
https://gardco.com/stormer_krebsconv_PU-G271.pdf; 4 pages. cited by
examiner .
Canadian Office Action, issued by the Canadian Intellectual
Property Office, regarding corresponding patent application Serial
No. CA 2,989,996; dated Jan. 17, 2019, 6 pages. cited by applicant
.
Auschra, C. et al., The Role of Thickeners in Optimizing Coatings
Formulation. [Retrieved from the internet Aug. 16, 2016]
<http://www.chinacoatcongress.net/con2014_CD/Paper_EN/Paper_7_BASF_E.p-
df>; 2014, p. 10, table 3 and second paragraph; p. 12, thrid
paragraph; p. 13, figure 15. cited by applicant .
Chroma-Chem, Products for Paint and Coatings. Apr. 2012; [Retrieved
from the internet Aug. 16, 2016]
,<http://www.pcimag.com/ext/resources/VirtualBrochureFeb2012/Chromaflo-
.pdf?1372101688>. p. 43, col. 1, first paragraph; p. 44 table.
cited by applicant .
Kim, M_S et al.; An In Vivo Study of the Host Tissue Response to
Subcutaneous Implantation of PLGA- and/or Porcine Small Intenstine
Submuscosa-based scaffolds. Biomaterials. vol. 28. Aug. 30, 2007;
p. 5138, col. 2, second paragraph. cited by applicant .
International Search Report and Written Opinion, issued by the
ISA/U.S. Receiving Office, regarding corresponding international
application Serial No. PCT/US2016/038088, dated Sep. 16, 2016, 18
pages. cited by applicant.
|
Primary Examiner: Klemanski; Helene
Attorney, Agent or Firm: Fox Rothschild LLP
Parent Case Text
CROSS REFERENCE TO RELATED APPLICATIONS
This application is a national stage patent application of
International patent application Serial No.: PCT/US2016/038088,
filed on Jun. 17, 2016; which claims the benefit of U.S.
Provisional patent application Ser. No. 62/182,153, filed on Jun.
19, 2015; the entireties of which are hereby incorporated by
reference.
Claims
We claim:
1. An ink composition for marking a tissue specimen comprising:
30.0 wt. % to 54.0 wt. % of an alkali soluble styrene; 0.3 wt. % to
1.7 wt. % cellulose; 8.0 wt. % to 48.0 wt. % colorant; 0.0% to 13.0
wt. % pigment; 23.0 wt. % to 47.0 wt. % solvent; and 0.35 wt. % to
1.65% of a defoamer, wherein said ink composition has a viscosity
of from 90 ku to 115 ku at room temperature and a reflective light
value in the L.a.b. color space in dimension L of from
approximately 31.4 to 84.51, in dimension a of from approximately
-42.32 to +52.81, and in dimension b of from approximately -30.73
to +88.09.
2. The ink composition of claim 1 further comprising 0.05 wt. % to
0.15 wt. % of an amine.
3. The ink composition of claim 2 wherein the amine is
amino-2-methyl-1-propanol 95% active (5% water).
4. The ink composition of claim 1 wherein the cellulose is selected
from ethyl hydroxyethyl cellulose and methyl ethyl hydroxyethyl
cellulose.
5. The ink composition of claim 4 wherein the cellulose is ethyl
hydroxyethyl cellulose.
6. The ink composition of claim 1 wherein the ink composition has a
peak transmission in the UV-visible light spectrum at a wavelength
of from 322 nm to 716 nm.
7. The ink composition of claim 1 further comprising a preservative
in an amount from 0.1 to 1.1 wt. %.
8. The ink composition of claim 1 wherein the alkali soluble
styrene is an acrylic styrene copolymer emulsion.
9. The ink composition of claim 7 wherein said preservative
comprises an amount from 0.2 to 1.0 wt. %.
10. The ink composition of claim 1 wherein the solvent comprises
deionized water.
11. The ink composition of claim 1 wherein the colorant comprises
from 7.59 to 11.59 wt. % carbazole violet and the pigment comprises
2.75% to 4.75 wt. % white mica.
12. The ink composition of claim 11 wherein said ink composition
has a peak transmission in the UV-visible light spectrum at a
wavelength of 451 nm.
13. The ink composition of claim 1 wherein the colorant comprises
from 24.48 to 28.48 wt. % phtahalo green yellow and the pigment
comprises 8.6 to 10.6 wt. % white mica.
14. The ink composition of claim 13 wherein said ink composition
has a peak transmission in the UV-visible light spectrum at a
wavelength of 545 nm.
15. The ink composition of claim 1 wherein the colorant comprises
from 21.28 to 25.28 wt. % phthalo blue green and the pigment
comprises 6.53 to 8.53 wt. % white mica.
16. The ink composition of claim 15 wherein said ink composition
has a peak transmission in the UV-visible light spectrum at a
wavelength of 506 nm.
17. The ink composition of claim 1 wherein the colorant comprises
from 15.18 to 19.18 wt. % naphthol red and the pigment comprises
4.40 to 8.40 wt. % quinacridone red.
18. The ink composition of claim 17 wherein said ink composition
has a peak transmission in the UV-visible light spectrum at
wavelengths of 449 nm and 716 nm.
19. The ink composition of claim 1 wherein the colorant comprises
from 4.22 to 8.22 wt. % benzimidazolone yellow, 4.16 to 8.16 wt. %
disazopyrazolone orange and the pigment comprises 1.5 to 3.5 wt. %
white mica.
20. The ink composition of claim 19 wherein said ink composition
has a peak transmission in the UV-visible light spectrum at
wavelengths of 352 nm and 636 nm.
21. The ink composition of claim 1 wherein the colorant comprises
from 25.63 to 29.63 wt. % benzimidazolone yellow.
22. The ink composition of claim 21 wherein said ink composition
has a peak transmission in the UV-visible light spectrum at
wavelengths of 322 nm and 605 nm.
23. The ink composition of claim 1 wherein said ink composition has
two peak transmissions in the UV-visible light spectrum at
wavelengths from 322 nm to 716 nm.
24. The ink composition of claim 1 wherein said ink composition has
a single peak transmission in the UV-visible light spectrum at a
wavelength from 451 nm to 545 nm.
25. The ink composition of claim 1 wherein said ink composition is
gamma irradiated with a minimum dose of 27.5 kGy up to a maximum
dose of 45 kGy.
26. The ink composition of claim 25 wherein said ink composition is
gamma irradiated with a minimum dose of 31.0 kGy to a maximum dose
of 39 kGy.
27. The ink composition of claim 26 wherein said ink composition is
gamma irradiated with a dose of 33 kGy.
28. The ink composition of claim 1 wherein the colorant comprises
from 9.79 wt. % to 13.79 wt. % benzimidazolone yellow and from 0.36
wt. % to 4.36 wt. % disazopyrazolone orange and from 5.72 wt. % to
9.72 wt. % orange.
29. The ink composition of claim 28 wherein said ink composition
has two peak transmission in the UV-visible light spectrum at
wavelengths from 475 nm to 675 nm.
30. An ink composition comprising: 30.0 wt. % to 54.0 wt. % of an
alkali soluble styrene; 0.3 wt. % to 1.7 wt. % cellulose; 8.0 wt. %
to 48.0 wt. % colorant; 0.0% to 13.0 wt. % pigment; 23.0 wt. % to
47.0 wt. % solvent; and 0.35 wt. % to 1.65% of a defoamer in
combination with a fixing solution having a composition of from
31.0-35.0% ethanol (95%), 10.1-12.1% lactic acid, 31.3-35.3%
deionized water and 21.0-23% formalin.
31. An ink composition for marking tissue comprising: 30.0 wt. % to
54.0 wt. % of an alkali soluble styrene; 0.3 wt. % to 1.7 wt. %
cellulose; from 15.6 wt. % to 19.60 wt. % lampblack colorant; 0.0%
to 13.0 wt. % pigment; 23.0 wt. % to 47.0 wt. % solvent; and 0.35
wt. % to 1.65% of a defoamer, wherein said ink composition has a
viscosity of from 90 ku to 115 ku at room temperature and a
reflective light value in the L.a.b. color space in dimension L of
25.81, in dimension a of -0.02, and in dimension b of -0.49.
32. The ink composition of claim 31 wherein said ink composition
has a lineal UV-Vis spectrum having no visible peak transmission
between 250 nm and 950 nm.
33. A method of preparing an ink composition for use in marking
tissue specimens comprising: preparing a first solution including
mixing from 0.3 wt. % to 1.7 wt. % ethyl hydroxyethyl cellulose
into 23.0 wt. % to 47.0 wt. % deionized water under agitation;
adding from 0.05 wt. % to 1.05 wt. % of amino-2-methyl-1-propanol
(95% active, 5% water); allowing said first solution to sit at room
temperature for 5 to 12 hours; preparing a final solution by mixing
from 30.0 to 54.0 wt. % alkali soluble styrene acrylic resin, 0.35
to 1.65 wt. % defoamer, 8.0 to 35.0 wt. % colorant, 0.0 to 13.0 wt.
% pigment to said first solution; at room temperature measuring a
viscosity of said final solution and adjusting said viscosity with
methyl ethyl hydroxyethyl cellulose until a desired range of from
90 ku to 115 ku is reached.
Description
FIELD OF THE INVENTION
The present invention relates generally to ink that is useful in
marking tissue specimens. In particular the invention relates to
one or more inks and a fixing solution that are used to mark tissue
specimens.
BACKGROUND OF THE INVENTION
When a surgeon removes cancerous tissue, it is carefully analyzed
to estimate if any cancerous cells remain in the patient's body.
The "tissue margin" is the edge or border of the tissue that has
been removed during surgery; specific "margins" may refer to a
subset of the exterior surface. A margin is designated as "clear"
when no cancer cells are found at the tissue's edge. Conversely, a
"positive" result is designated when cancer cells are found at the
edge of the tissue, and the implication is that not all cancer was
removed during surgery. In these cases, a second surgery or some
other type of clinical treatment to address the remaining cancer
cells may be recommended. Tissue specimens are evaluated in the
pathology lab after surgery. For some types of surgery, such as
breast cancer surgery, the excised tissue specimen is also
evaluated by X-ray during the operation. The clinical status of the
tissue margins is considered one of the most important factors in
predicting whether there will be a recurrence of the cancer. The
accurate identification and reporting of the clinical status of
tissue margins (i.e., "clear of cancerous cells" or "positive for
cancerous cells") is clinically relevant in a wide range of
pathology specimens. Tissue marking inks are an important tool used
in the determination of margin status and thereby affect the
subsequent clinical action taken. Depending on the type of surgery,
different colors of inks are used on a single specimen to
designate: the specimen exterior surface margin; specific "margins"
(or areas that are a subset of the entire specimen surface);
particular anatomical features, or particular areas of concern on
the specimen. Sometimes a "fixing solution" is applied after the
inks to strengthen the adhesion of the ink to the tissue. Either
before or after the ink is applied, the tissue is typically
preserved in a "fixative" such as formalin. If the pathology
analysis has a "positive" finding, the report informs the physician
of the location in the patient's body where additional tissue
should be removed or additional treatment directed.
A tissue specimen is often an irregular-shaped piece of tissue with
fissures, crevasses or flaps on the surface. The tissue needs to be
marked or labeled to designate the specimen margin(s) and original
position of the tissue in the patient's body. The method of marking
the tissue, which is typically to use ink, must mark accurately
despite the irregularities in the surface of the specimen. If
pathology analysis shows that cancerous cells are close to the
exterior surface of the specimen, there may be additional cancerous
cells remaining in the patient's body, requiring a subsequent
surgery or other clinical treatment. For this subsequent surgery or
clinical treatment to effectively address the any remaining cancer
cells, it is essential that the original shape and location of the
excised tissue is accurately determined. Inks that label the
anatomy or the positioning of the excised tissue to indicate how it
is originally positioned in the patient's body provide direction on
where to target the subsequent surgery or treatment in order to
more completely eradicate the cancer. Pathologists typically use
inks to mark the exterior surfaces, or margins of excised tissue in
cancer surgery. In some cases, the surgeon marks excised tissue
before it is sent to the pathology lab.
Inks must adhere to tissue effectively for an accurate analysis in
the pathology lab. They must dry quickly when applied to the
tissue, both for the efficiency of use and also to maintain the
fidelity of the markings without smearing or migrating on the
tissue surface. The pathology lab makes slides of microscopic
tissue specimens for analysis. Various materials and procedures are
used to "process" the tissue specimen. The tissue is fixed, often
using formalin; the pathologist then takes a slice of the tissue,
typically approximately 5 microns thick, and places it on a glass
slide. The slides are evaluated to identify the presence of
cancerous cells, and the distance from the cancerous cells to the
"margin," or the exterior of the specimen, is measured. A wider
distance indicates that the cancerous cells are less likely to have
spread beyond the boundaries of the excised tissue. A shorter
distance suggests that there may be cancerous cells remaining in
the patient's body, and a surgical re-excision or other clinical
treatment to eradicate any remaining cancer is often performed. The
success of this subsequent surgery or treatment may depend upon an
accurate understanding of how the specimen was originally shaped
and positioned in the patient's body, or the accurate
identification of important anatomical features on the specimen.
The location of the cancer cells that are close to the outer
surface of the specimen or on a particular anatomical feature
indicate the corresponding locations where remaining cancer cells
may be present in the patient. Thus, it is critical that the ink
that is applied to the surface of the tissue dries quickly and
adheres securely, maintaining the integrity of the marking, without
running, dripping or migrating to an adjacent area on the surface
of the specimen or into a fissure or crevasse. If the ink does not
dry quickly the fidelity of the markings are affected and the ink
may smear or migrate on the tissue surface. If the ink migrates to
an adjacent area on the surface of the tissue, and that area is
determined to have cancerous cells close to the surface, then the
surgical re-excision or clinical treatment directed at remaining
cells in the patient may target the wrong location, resulting in
untreated cancer cells which may cause a recurrence of the cancer.
The most serious negative outcome is a local recurrence of the
cancer. Of the 4% to 20% of lumpectomy patients who suffer a
recurrence of their breast cancer, half of these cases are
metastatic, or potentially deadly. If the ink migrates into a
fissure or crevasse in the tissue, the pathology report may result
in a "false positive," because the cancerous cells appear closer to
the exterior surface of the tissue specimen than they actually
were. A false positive could result in unnecessary surgery or
clinical treatment for the patient. In breast cancer cases,
re-excisions occur for up to 60% of lumpectomy patients, and of
these, as many as 66% are false positives.
The viscosity of commercially available inks is widely variable.
Thin inks often run and drip when applied to tissue, which prevents
the accurate designation of specific tissue margins, which is the
purpose for inking tissue. Moreover, thin inks have a great
percentage of water and take a long time to dry. Thin inks can also
run into crevasses of tissue, whereby the ink becomes closer to the
cancer cells in the specimen, causing the margins to be
misinterpreted, and creating a false positive result that may cause
an unnecessary second surgery or treatment. On the other hand,
thick inks can be lumpy and sometimes contain dry chards of
crystallized matter, which appear to be microcalcifications
(potentially cancerous cells) on the X-ray image taken during the
surgical procedure. These dry fragments of ink may cause false
positives in the interpretation of the X-ray image, resulting in
unnecessary removal of additional tissue, or at a minimum causing
confusion and delay during the operation while the patient is
anesthetized.
Adherence to tissue can be problematic with commercially available
inks when these inks are applied to tissue and the tissue is fixed
in formalin. Tissue may be fixed in formalin before or after ink is
applied. If tissue is fixed in formalin first, then the inks often
adhere less effectively to the tissue. Commercially available inks
do not adhere well to tissue that has been pre-fixed in formalin.
If the inks are applied first and then the inked specimen is
submerged into formalin, the inks may wash off of the tissue and
into the formalin.
Commercially available inks are not sterile, which also results in
problems. First, non-sterile inks may allow cross-contamination
among tissue specimens because the applicator used to apply the ink
may touch the specimen and then touch the bottle during the process
of applying ink. Second, if the surgeon applies non-sterile inks
while in the operating room, the non-sterile inks may not be used
in the sterile field, which introduces the risk of errors if the
specimen is carried across the room to the non-sterile area before
it is inked or if inking is delayed until the surgery is complete.
Third, non-sterile inks sometimes feature dry fragments that look
like microcalcifications, or potentially cancerous cells, on the
intraoperative X-ray image.
Other problems exist with commercially available inks. Although
accurate identification of ink color is essential to effective
clinical usage of the inks, the colors of commercially available
inks are often difficult to distinguish from one another. To
perform effectively, each ink color must be both recognizable and
distinguishable from other colors under both reflective light
(ordinary lighting conditions, as when the ink is applied to tissue
in the pathology lab or operating room), and under transmitted
light (when light shines through from the opposite side, as when
the tissue on a slide is placed under a microscope). When the inks
are viewed under reflective light, dark colors such as blue, violet
and green are often difficult to distinguish from one another and
all appear to be black. When applied to tissue and viewed on the
slides under a microscope using transmitted light, difficulties are
often encountered with the yellow-orange, orange-red, blue-violet
and red-violet comparisons. Confusion accurately identifying the
ink color on the specimen can lead to erroneous interpretation
regarding the location of cells that are close to the specimen
surface, resulting in re-excision or treatment in an incorrect
location, and possibly cancer recurrence.
When inks are applied to tissue specimens, a fixing solution is
sometimes used. The purpose of the fixing solution is to enhance
the adherence of the inks to the tissue. However, commercially
available fixing solutions are problematic in that they fail to
effectively strengthen the adherence of the inks to the tissue,
resulting in ink washing off when tissue is pre-treated with
formalin before inking, submerged in formalin after ink is applied,
or when a knife is used to cut through the inked specimen. In
addition, some fixing solutions have an odor that is offensive to
clinical staff.
Accordingly, there is a need for colored inks for marking tissue
specimens that overcome the foregoing problems of commercially
available inks. There is also a need for a fixing solution that
prevents the wash-off of inks from the tissue specimens when the
tissue is exposed to formalin before or after ink is applied.
BRIEF SUMMARY OF THE INVENTION
The problems outlined above are addressed by the ink for marking
tissue specimens in accordance with the invention. The inks in
accordance with the invention adhere well to a wide variety of
tissue types, do not run, drip, bleed or smear onto adjacent tissue
margins or into the interior of the specimen when the tissue is
cut; adhere to tissue that has been previously fixed in formalin
before ink is applied; adhere to tissue when the specimen is placed
into formalin after inks are applied; have colors that are
recognizable and distinct from other colors under both reflective
light and transmitted light; are not detectable on an X-ray and do
not leave artifacts that are visible on imaging, dry when applied
to tissue within 2-3 minutes or less, and maintain color and
performance characteristics when exposed to gamma radiation.
The foregoing is accomplished by ink compositions having a general
formula of 30.0 wt. % to 54.0 wt. % of an alkali soluble styrene;
0.3 wt. % to 1.7 wt. % ethyl hydroxyethyl cellulose; 8.0 wt. % to
35.0 wt. % colorant; 0.0 wt. % to 13.0 wt. % pigment; 23.0 wt. % to
47.0 wt. % deionized water; 0.35 wt. % to 1.65% defoamer and 0.1
wt. % to 1.1 wt. % preservative. The inks in accordance with the
invention may have a peak transmission in the visible spectrum at a
wavelength of from 322 nm to 716 nm or having a lineal UV-Vis
spectrum with no visible peak transmission between 250 nm and 950
nm.
Inks in accordance with the invention may be utilized to identify
the orientation of a tissue specimen. The method of identifying the
orientation of a tissue specimen having first, second and third
surfaces includes providing at least three inks having three
different colors each having a composition of 30.0 wt. % to 54.0
wt. % of an alkali soluble styrene; 0.3 wt. % to 1.7 wt. % ethyl
hydroxyethyl cellulose; 8.0 wt. % to 35.0.0 wt. % colorant; 0.0 wt.
% to 13.0 wt. % pigment; 23.0 wt. % to 47.0 wt. % deionized water;
0.35 wt. % to 1.65% defoamer and 0.1 wt. % to 1.1 wt. %
preservative, said ink composition having properties that causes
each of said at least three inks to adhere to the first, second and
third surfaces of a tissue specimen, respectively, without bleeding
into a crevice of the tissue specimen or onto an adjacent tissue
margin.
The inks are also used with a novel fixing solution that overcomes
the problems with conventional fixing solutions. The fixing
solution has a formulation of from 31.0-35.0% ethanol (95%),
10.1-12.1% lactic acid, 31.3-35.3% deionized water and 21.0-23%
formalin.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is a front or anterior view of a patient including a tissue
specimen to be removed from the patient.
FIG. 2 is an enlarged front or anterior view of the portion of the
patient and tissue specimen indicated at circle 2 of FIG. 1.
FIG. 3 is a side or lateral view of the portion of the patient and
tissue specimen indicated at lines 3-3 of FIG. 2.
FIG. 4 is a perspective view of the tissue specimen of FIG. 1 after
being removed from the patient of FIG. 1 showing three sides of the
tissue that will be marked, in this case, superior, medial and
lateral.
FIG. 5 is a graph from 400-800 nm depicting the light transmission
characteristics of conventional inks.
FIG. 6A is a graph from 220-920 nm depicting the light transmission
characteristics of various ink composition in accordance with the
invention.
FIG. 6B is a graph from 250-950 nm depicting the light transmission
characteristics of the black ink composition in accordance with the
invention.
FIGS. 7 and 8 are a visual depiction of how a commercially
available ink drips and run into other adjacent ink when applied on
tissue.
FIG. 9A is a radiograph of a tissue specimen depicting the
histologic appearance of actual microcalcifications in the
tissue.
FIG. 9B is a radiograph of a tissue specimen marked with
commercially available ink showing the false appearance of a
cluster of microcalcifications.
FIG. 10 is an X-ray of tissue marked with the ink in accordance
with the present invention showing no appearance of
calcifications.
DETAILED DESCRIPTION OF THE INVENTION
As used herein the following terms have the definitions set forth
next to them.
"Pigment" means the dry powder used to prepare and contained in a
colorant or ink.
"Colorant" means a pigment dispersed in water with acrylic resin
added.
"Ink" and "Ink Composition" means a colorant with the additional
ingredients set forth herein, which is in final form and ready to
be applied to tissue.
"Fixing solution" according to the invention described herein means
a liquid mixture having a composition of from 31.0-35.0% ethanol
(95%), 10.1-12.1% lactic acid, 31.3-35.3% deionized water and
21.0-23% formalin. The fixing solution enhances the adherence of
the inks to the tissue.
"Fixative" means a colorless solution of formaldehyde in water
(formalin) used to preserve biological specimens. Alternative
"fixatives" include Weigners, Bouins, Hollandes, GreenFix, UPM,
Cymol, Excell-Plus, FineFix, RCL2, HOPE, Glyo-Fixx, Cell-block.
With reference to FIGS. 1-3, a patient 10 and a tissue specimen 15
are illustrated to show the tissue specimen's orientation in the
patient 10. Before proceeding, it should be noted that the present
invention will be described as it relates to a tissue specimen 15
removed from a breast. However, one of ordinary skill in the art
will realize that the invention is applicable to many other types
of tissue specimens in which tissue margin identification is
important. In addition, those of skill in the art will appreciate
that the inks and fixing solution in accordance with the invention
may be used in both human subjects and animals.
For example, pancreatic cancer requires that a tissue specimen be
removed, and that the original orientation and anatomical features
be identified. As such, the invention should not be limited only to
the uses described herein as it is well suited for use with any
excised tissue that requires for pathology analysis designation of
the specimen's: original orientation, the exterior margin surface
or subsets of that surface, specific anatomical features, or
particular areas of concern. These tissues include but are not
limited to specimens of breast, pancreas, bone, thyroid, lymph
nodes, brain, sarcomas, kidney, bowel, spleen, soft tissue masses,
melanoma, squamous cell skin cancer, basal cell cancer, liver
tumors, and the like.
FIG. 1 illustrates how the inks may be applied to a specimen to
indicate the original orientation, or position, in the patient 10.
For purposes of description, the direction 20 toward the patient's
head will be identified as superior, while the opposite direction
25 is inferior. The direction 30 toward the patient's midline is
defined as medial, while the opposite direction 35 is defined as
lateral. With reference to FIG. 3, a side or medial view of a
portion of the patient is shown to further illustrate orientation.
The direction 40 toward the patient's front exterior is defined as
anterior, while the opposite direction 45 is defined as
posterior.
FIGS. 2 and 3 illustrate the tissue specimen 15 within the patient
10 prior to its removal, while FIG. 4 shows that same tissue
specimen 15 after removal. With the specimen 15 still in the
patient 10, the three surfaces 50, 55, 60 that will be marked can
be seen. While any three planes or surfaces of the tissue specimen
15 can be used to identify the orientation of the specimen 15, it
is preferred that at least three substantially orthogonal surfaces
be identified, with some applications marking six or more surfaces.
In FIGS. 2 and 3, the lateral surface 50, medial surface 55, and
superior surface 60 of the tissue specimen 15 are shaded
differently for illustrative purposes. In FIG. 4, the same three
surfaces 50, 55, 60 are shaded to indicate that each one is marked
with a different color.
Typically, to mark the surfaces 50, 55, 60, three different colored
inks are employed. Ink colors in accordance with the invention
include violet, green, blue, red, orange and yellow. However, those
of skill in the art will appreciate that any color ink is
contemplated to be within the scope of the invention so long as it
meets the general formulation set forth in Table I.
Preferred ranges for the ink compositions in accordance with the
invention are set forth in the Tables below. The inks in accordance
with the invention have a viscosity of from 90 Krebs units (ku) to
115 ku and more preferably between 95 ku and 110 ku.
TABLE-US-00001 TABLE I GENERAL INK FORMULATION Ingredient Most
Preferred Range Preferred Range Alkali soluble styrene 32.0-52.0
wt. % 30.0-54.0 wt. % acrylic resin Cellulose 0.5-1.5 wt. % 0.3-1.7
wt. % Colorant (dispersed 10.0-33.0 wt. % 8.0-35.0 wt. % pigment)
(available from Chromoflow Technologies, Somerset, NJ) Pigment
White Mica 6300 0.0-12.0 wt. % 0.0-13.0 wt. % (available from A.E.
Fleming, Warren, MI) Deionized water 25.0-45.0 wt. % 23.0-47.0 wt.
% Amine 0.05%-0.10% wt. % 0.05-0.15 wt. % Defoamer (BYK-011,
0.5-1.5 wt. % 0.35-1.65 wt. % BYK-012 or both) available from
Altana AG: Wesel, Germany) Preservative 0.2-1.0 wt. % 0.1-1.1 wt.
%
The ranges set forth above are critical to the inks in accordance
with the invention. For example with respect to alkali soluble
styrene acrylic resin, a composition having less than 30 wt. % will
result in an ink that forms a brittle film on tissue with poor
adhesion. A resin composition of more than 54.0 wt. % will result
in an ink having poor or inadequate opacity, resulting in less
visibility when applied to the tissue or on the slides. The
inventors have found that in the claimed ink composition resin is
one of the keys to superior adherence to tissue and fast drying on
tissue, and is moisture resistant in an acidic atmosphere such as
formalin. Notably, this composition is effective on fatty tissue,
which is one of the most difficult tissue types on which to achieve
adhesion. Examples of water-dispersible resins include synthetic
resins such as polyester resins, polyurethane resins, polyepoxy
resins, polyamide resins, polyether resins, poly(meth)acrylic
resins, acryl-silicone resins, fluorine-based resins, polyolefin
resins, polystyrene-based resins, polybutadiene-based resins,
polyvinyl acetate-based resins, polyvinyl alcohol-based resins,
polyvinyl ester-based resins, polyvinyl chloride-based resins,
polyacrylic acid-based resins, unsaturated carboxylic acid-based
resins and copolymers such as styrene-acrylate copolymer resins,
styrene-butadiene copolymer resins, and combinations of the
plural.
Preferred resins may be selected from DSM Neoresins, e.g. the
NeoCryl product line, in particular acrylic styrene copolymer
emulsions NeoCryl A-662, NeoCryl A-1131, NeoCryl A-2091, NeoCryl
A-550, NeoCryl BT-101, NeoCryl SR-270, NeoCryl XK-52, NeoCryl
XK-39, NeoCryl A-1044, NeoCryl A-1049, NeoCryl A-1110, NeoCryl
A-1120, NeoCryl A-1127, NeoCryl A-2092, NeoCryl A-2099, NeoCryl
A-308, NeoCryl A-45, NeoCryl A-615, NeoCryl BT-24, NeoCryl BT-26,
NeoCryl BT-26, NeoCryl XK-15, NeoCryl X-151, NeoCryl XK-232,
NeoCryl XK-234, NeoCryl XK-237, NeoCryl XK-238-NeoCryl XK-86,
NeoCryl XK-90 and NeoCryl XK-95. Preferably NeoCryl A-2092 is
used.
Similarly, an ink having a composition of less than 0.3 wt. % of
cellulose will result in an ink having a low viscosity which in
turn will lead to running and dripping on the tissue surface and
migration into tissue crevices, resulting in inaccurate
identification of tissue margins. Similarly, an ink having a
cellulose composition of more than 1.7 wt. % will result in an ink
having a viscosity higher than desirable, resulting in poor
"transfer" capability. Poor transfer occurs when the ink is
difficult to move in the process of loading it onto an applicator
and then releasing it onto the tissue; adhesion to the tissue is
problematic as well. Cellulose may be selected from ethyl
hydroxyethyl cellulose or methyl ethyl hydroxyethyl cellulose and
is preferably ethyl hydroxyethyl cellulose available as Bermocoll
from Akzo Nobel N.V.: NL.
If the amount of colorant is less than 8.0 wt. % then the result is
poor color differentiation when compared to other colors and poor
correspondence between a particular color when viewed under
reflective versus transmitted light. Conversely, if the amount of
colorant is greater than 35.0 wt. % the resulting ink is brittle
(when applied to tissue), resulting in distortion of the color
under both reflective and transmitted light. In addition, the ink
will have poor adherence to the tissue.
Water is an environmentally friendly and hence desirable solvent.
In the present invention, the content of water to the whole ink
composition is set forth in the Table above. An ink composition
having less than 23.0 wt. % of water will be highly viscous and
have poor transfer from the ink applicator to tissue. A water
composition of more than 47 wt. % will result in an ink that is
watery and has too low a viscosity, resulting in dripping, running,
and migration when applied to tissue, resulting in the loss of
fidelity between where the ink is applied and the ultimate
interpretation of the margins. An additional result is poor "film
build," or the thickness with which a layer of ink adheres to the
tissue; poor film build compromises opacity, or the vividness with
which the ink color is visible on the tissue.
Amine is added to the formula to help the cellulosic thickener
activate and to adjust the pH to the desired range of approximately
pH 8.5. If an ink has too little amine the resin will gel and lose
its water solubility. If an ink composition has greater than 0.15
wt. % of amine, the pH of the system is too basic and it requires
additional amounts of the fixing solution with an acidic pH to
bring the pH below 7.0 so that the resin becomes insoluble. The
insolubility of the resin is needed so that the ink does not bleed
into the formalin solution.
With respect to a defoamer, an ink composition having less than
0.35 wt. % would be susceptible to foaming and lack viscosity
stability, resulting in an apparent change in volume or density
over time as the finished product is handled, shipped, or stored.
For example, ink may completely fill a container upon packaging,
but when the container is opened at a later date, the volume of ink
may appear significantly reduced. An ink composition having a
defoamer component greater than 1.65 wt. % would have
irregularities (holes or weak spots in the coverage) because
surface tension of the ink is too low. This may diminish the
readability of the inks on the microscopic slides.
A preservative is used to prevent bacteria or mold growth in the
inks. Preferable preservatives may be selected from the Euxyl line
of products and include phenoxyethanol and ethylhexylglycerin
available from Scheulke and Mayr (Norderstedt, Germany). A most
preferable preservative is Euxyl PE-9010. The minimum amount of a
preservative that is necessary is 0.1 wt. %. Consequences of an
inadequate amount of preservative include bacteria and/or mold
growth in the ink, which can distort the color under both
reflective and transmitted light. For example, mold may appear as
black spots in the ink under both reflective and transmitted light.
Consequences of exceeding 1.1 wt. % of a preservative is an ink
that has a low surface tension and uneven coverage on tissue.
In the ink composition of the present invention, a
water-dispersible colorant is present. The ink composition may
include one or more colorants and a pigment. It is critical that
the colorants used in the inks in accordance with the invention be
compatible with the resin system. In addition, each ink must be
bright, recognizable, and easily distinguished from each different
color of ink under both reflective and transmitted light,
especially violet, green and black. Other critical ink
characteristics include gloss and color "stability;" stability
refers to the persistence of the color when exposed to acids or
alkalis. Thus, the particular colorants selected for the inks in
accordance with the invention achieve a balance between providing
sufficient opacity to be distinguished visually when illuminated by
both reflective light and transmitted light at various
magnifications and exposures. Thus, the ink compositions in
accordance with the invention have peak transmissions in the UV and
visible spectrum at the wavelengths best seen FIGS. 5-6. White mica
6300 in amounts from 0.0-13.0 wt. % were added to the ink
compositions to achieve the required brightness and color
differentiation under reflected light. White mica 6300 increases
the brightness of the colors under reflective light without
occluding the transmitted light when viewed under the microscope.
One may consider using TiO2 as a brightener for reflective light;
however, the inventors have found that TiO2 occludes the passage of
light and creates a shadow, obscuring the ink color when viewed
under transmitted light. Acceptable colorants are selected from the
Plasticolors UCD-E line as noted below.
TABLE-US-00002 TABLE II Color Pigment - Colorants Violet White Mica
6300 UCD 8406E Carbazole Violet (R-175) Green White Mica 6300 UCD
5166E Phtahalo Green Yellow Shade (G-36) Blue White Mica 6300 UCD
4820E Phthalo Blue Green Shade (BL 15-3) Red UCD 7949E Naphthol Red
(BS - R-170) plus UCD 8030E Quinacridone Red (V-19) Orange White
Mica 6300 UCD 5696E Benzimidazolone Yellow (Y-151) plus UCD 6012E
Disazopyrazolone Orange (O-34) Orange B None UCD 5696E
Benzimidazolone Yellow (Y- 151) plus UCD 6012E Disazopyrazolone
Orange (O-34) plus UCD 6002E Orange 43 Yellow None UCD 5696E
Benzimidazolone Yellow (Y-151) Black None UCD 1625E Lampblack
(BK-7)
Preferred ranges for the ink compositions in accordance with the
invention are set forth in the Tables below. The color referenced
below refers to the colorants identified in TABLE II above.
TABLE-US-00003 TABLE III VIOLET Most Ingredient Preferable More
Preferable Preferable Resin A2902 47.51 wt. % 47.01 wt. % to 48.01
wt. % 45.51 wt. % to 49.51 wt. % Thickener Cellulose 1.00 wt. %
0.90 wt. % to 1.10 wt. % 0.80 wt. % to 1.20 wt. % Colorant Violet
9.59 wt. % 9.39 wt. % to 9.79 wt. % 7.59 wt. % to 11.59 wt. %
Pigment White Mica 3.75 wt. % 3.55 wt. % to 3.95 wt. % 2.75 wt. %
to 4.75 wt. % 6300 Water Deionized 37.60 wt. % 36.10 wt. % to 39.10
wt. % 35.60 wt. % to 39.60 wt. % Amine AMP 95 0.05 wt. % 0.05 wt. %
to 0.10 wt. % 0.05 wt. % to 0.15 wt. % Defoamers 0.90 wt. % 0.80
wt. % to 1.00 wt. % 0.75 wt. % to 1.05 wt. % Preservative 0.50 wt.
% 0.45 wt. % to 0.55 wt. % 0.40 wt. % to 0.60 wt. %
TABLE-US-00004 TABLE IV GREEN Most Ingredient Preferable More
Preferable Preferable Resin A2902 34.83 wt. % 34.33 wt. % to 35.33
wt. % 32.83 wt. % to 36.83 wt. % Thickener Cellulose 0.70 wt. %
0.60 wt. % to 0.80 wt. % 0.50 wt. % to 0.90 wt. % Colorant Green
26.48 wt. % 25.98 wt. % to 26.98 wt. % 24.48 wt. % to 28.48 wt. %
Pigment White 9.60 wt. % 9.10 wt. % to 10.10 wt. % 8.60 wt. % to
10.60 wt. % Mica 6300 Water Deionized 27.06 wt. % 25.56 wt. % to
28.56 wt. % 25.06 wt. % to 29.06 wt. % Amine AMP 95 0.05 wt. % 0.05
wt. % to 0.10 wt. % 0.05 wt. % to 0.15 wt. % Defoamer 0.90 wt. %
0.80 wt. % to 1.0 wt. % 0.75 wt. % to 1.05 wt. % Preservative 0.50
wt. % 0.45 wt. % to 0.55 wt. % 0.40 wt. % to 0.60 wt. %
TABLE-US-00005 TABLE V BLUE Most Ingredient Preferable More
Preferable Preferable Resin A2902 37.61 wt. % 37.11 wt. % to 38.11
wt. % 35.61 wt. % to 39.61 wt. % Thickener Cellulose 0.75 wt. %
0.65 wt. % to 0.85 wt. % 0.55 wt. % to 0.95 wt. % Colorant Blue
23.28 wt. % 22.78 wt. % to 23.78 wt. % 21.28 wt. % to 25.28 wt. %
Pigment White 7.53 wt. % 7.03 wt. % to 8.03 wt. % 6.53 wt. % to
8.53 wt. % Mica 6300 Water Deionized 29.57 wt. % 28.07 wt. % to
31.07 wt. % 27.57 wt. % to 31.57 wt. % Amine AMP 95 0.05 wt. % 0.05
wt. % to 0.10 wt. % 0.05 wt. % to 0.15 wt. % Defoamers 0.83 wt. %
0.73 wt. % to 0.93 wt. % 0.68 wt. % to 0.98 wt. % Preservative 0.45
wt. % 0.40 wt. % to 0.50 wt. % 0.35 wt. % to 0.55 wt. %
TABLE-US-00006 TABLE VI RED Most Ingredient Preferable More
Preferable Preferable Resin A2902 41.60 wt. % 41.10 wt. % to 42.10
wt. % 39.60 wt. % to 43.60 wt. % Thickener Cellulose 0.83 wt. %
0.73 wt. % to 0.93 wt. % 0.63 wt. % to 1.03 wt. % Colorant UCD
17.18 wt. % 16.83 wt. % to 17.53 wt. % 15.18 wt. % to 19.18 wt. %
7949E Naphthol Red (BS-R-170) Colorant UCD 6.40 wt. % 6.30 wt. % to
6.50 wt. % 4.40 wt. % to 8.40 wt. % 8030E Quinacridone Red (V-19)
Water Deionized 32.70 wt. % 31.20 wt. % to 34.20 wt. % 30.70 wt. %
to 34.70 wt. % Amine AMP 95 0.05 wt. % 0.05 wt. % to 0.10 wt. %
0.05 wt. % to 0.15 wt. % Defoamers 0.91 wt. % 0.81 wt. % to 1.01
wt. % 0.76 wt. % to 1.06 wt. % Preservative 0.45 wt. % 0.40 wt. %
to 0.50 wt. % 0.35 wt. % to 0.55 wt. %
TABLE-US-00007 TABLE VII ORANGE Most Ingredient Preferable More
Preferable Preferable Resin A2902 46.64 wt. % 46.14 wt. % to 47.14
wt. % 44.64 wt. % to 48.64 wt. % Thickener Cellulose 0.96 wt. %
0.86 wt. % to 1.06 wt. % 0.76 wt. % to 1.16 wt. % Colorant UCD
5696E 6.22 wt. % 6.02 wt. % to 6.42 wt. % 4.22 wt. % to 8.22 wt. %
Benzimidazolone Yellow (Y-151) Colorant UCD 6012E 6.16 wt. % 5.96
wt. % to 6.36 wt. % 4.16 wt. % to 8.16 wt. % Disazopyrazolone
Orange (O-34) Pigment White 2.50 wt. % 2.30 wt. % to 2.70 wt. %
1.50 wt. % to 3.50 wt. % Mica 6300 Water Deionized 35.96 wt. %
34.46 wt. % to 37.46 wt. % 33.96 wt. % to 37.96 wt. % Amine AMP 95
0.05 wt. % 0.05 wt. % to 0.10 wt. % 0.05 wt. % to 0.15 wt. %
Defoamers 1.05 wt. % 0.95 wt. % to 1.15 wt. % 0.90 wt. % to 1.20
wt. % Preservative 0.53 wt. % 0.48 wt. % to 0.58 wt. % 0.43 wt. %
to 0.63 wt. %
TABLE-US-00008 TABLE VIII ORANGE B Most Ingredient Preferable More
Preferable Preferable Resin A2902 42.95 wt. % 42.45 wt. % to 43.45
wt. % 40.95 wt. % to 44.95 wt. % Thickener Cellulose 0.85 wt. %
0.75 wt. % to 0.95 wt. % 0.65 wt. % to 1.05 wt. % Colorant UCD
5696E 11.79 wt. % 11.59 wt. % to 11.99 wt. % 9.79 wt. % to 13.79
wt. % Benzimidazolone Yellow (Y-151) Colorant UCD 6012E 2.36 wt. %
2.16 wt. % to 2.56 wt. % 0.36 wt. % to 4.36 wt. % Disazopyrazolone
Orange (O-34) Colorant UCD 6002E 7.72 wt. % 7.52 wt. % to 7.92 wt.
% 5.72 wt. % to 9.72 wt. % Orange 43 Water Deionized 33.05 wt. %
31.55 wt. % to 34.55 wt. % 31.05 wt. % to 35.05 wt. % Amine AMP 95
0.05 wt. % 0.05 wt. % to 0.10 wt. % 0.05 wt. % to 0.15 wt. %
Defoamers 0.65 wt. % 0.55 wt. % to 0.75 wt. % 0.50 wt. % to 0.80
wt. % Preservative 0.55 wt. % 0.50 wt. % to 0.60 wt. % 0.45 wt. %
to 0.65 wt. %
TABLE-US-00009 TABLE IX YELLOW Most Ingredient Preferable More
Preferable Preferable Resin A2902 39.45 wt. % 38.95 wt. % to 39.95
wt. % 37.45 wt. % to 41.45 wt. % Thickener Cellulose 0.78 wt. %
0.68 wt. % to 0.88 wt. % 0.58 wt. % to 0.98 wt. % Colorant Yellow
27.63 wt. % 27.13 wt. % to 28.13 wt. % 25.63 wt. % to 29.63 wt. %
Water Deionized 31.01 wt. % 29.51 wt. % to 32.51 wt. % 29.01 wt. %
to 33.01 wt. % Amine AMP 95 0.05 wt. % 0.05 wt. % to 0.10 wt. %
0.05 wt. % to 0.15 wt. % Defoamer 0.96 wt. % 0.86 wt. % to 1.06 wt.
% 0.81 wt. % to 1.11 wt. % Preservative 0.49 wt. % 0.44 wt. % to
0.54 wt. % 0.39 wt. % to 0.59 wt. %
TABLE-US-00010 TABLE X BLACK Ingredient Most Preferable More
Preferable Preferable Resin A2902 44.88% 44.38% to 45.38% 42.88% to
46.88% Thickener Cellulose 0.90% 0.80% to 1.00% 0.70% to 1.10%
Colorant Black 17.60% 17.35% to 17.85% 15.60% to 19.60% Water
Deionized 36.03% 34.53% to 37.53% 34.03% to 38.03% Amine AMP 95
0.05% 0.00% to 0.10% 0.00% to 0.10% Defoamers 0.99% 0.89% to 1.09%
0.84% to 1.14% Preservative 0.49% 0.44% to 0.54% 0.39% to 0.59%
The following examples illustrate the preparation of the ink
compositions in accordance with the invention.
Preparation of Ink Compositions
Example A
The method of preparing the ink compositions includes mixing two
solutions, A and B. Solution A (Cellulose Thickener) is prepared
using from 23.0 wt. % to 47.0 wt. % of deionized water and 0.3 wt.
% to 1.7 wt. % of ethyl hydroxyethyl cellulose (Bermocoll EBM
5500--Akzo Nobel). The deionized water is added to a mixing vessel
and the cellulose is gradually added to the water under agitation.
A dispersion type blade, such as a Cowels disperser, is used for
mixing. This solution can be mixed for approximately 20 to 35
minutes until smooth and lump free. From 0.05 wt. % to 1.05 wt. %
of amino-2-methyl-1-propanol 95% active (5% water) (AMP 95--Angus
Chemical) is then added and mixed into solution. After the solution
thickens it is mixed further for 20-30 minutes. Mixing speed is
increased as the thickening occurs. The speed of mixing is adjusted
so that a vortex occurs in the solution. The solution is covered
and allowed to sit for 5-12 hrs. or overnight at room temperature
before mixing with Solution B.
Solution B, the final ink composition is then formed using the
following materials:
TABLE-US-00011 NeoCryl A 2092 Solution 30.0-54.0 wt. % Colorants
8.0-35.0 wt. % Mica 0-13 wt. % Defoamer 0.35-1.65 wt. %
Preservative 0.1-1.0 wt. %
NeoCryl A 2092, the defoamers and Solution A, from above, are added
to a mixing vessel and mixed until smooth and lump free with a
dispersion blade. Colorant and a preservative are added under
agitation while blending with a Cowels disperser. Mica may also be
added depending on the ink composition. The solution is mixed for
10-15 minutes and the viscosity is checked. To achieve the desired
viscosity, additional methyl ethyl hydroxyethyl cellulose
(available as Bermocoll 5500 from AkzoNobel) may be added to the
vortex under vigorous agitation until the viscosity of the solution
reached is preferably between 90 Krebs units (ku) to 115 ku and
more preferably between 95 ku and 110 ku. Viscosities are measured
using a Brookfield KU-1 Viscometer. The solution is continued to be
mixed for a minimum of an additional 20 minutes and the viscosity
is measured again. When the desired viscosity of from 90 Krebs
units to 115 Krebs units is reached, the solution is allowed to
cool to room temperature. The resulting solutions are measured on
an Agilant 8451 UV-Vis spectrophotometer and diluted until the
violet, green, blue, red, orange, orange B and yellow ink
compositions have a peak transmissions of from 322 nm to 716
nm.
Example B
A violet ink composition is prepared. Solution A (Thickener) is
prepared using 35.60 wt. % of deionized water and 1.2 wt. % of
ethyl hydroxyethyl cellulose (Bermocoll EBM 5500--Akzo Nobel). The
deionized water is added to a mixing vessel and the cellulose is
gradually added to the water under agitation. A Cowels dispersion
blade is used for mixing. The solution is mixed for 25 minutes
until smooth and lump free. AMP 95 in an amount of 0.10 wt. % is
then added and mixed into solution. After initial thickening of the
solution, it is further mixed for 30 minutes, increasing the mixing
speed as the thickening occurred. The speed of mixing is adjusted
so that a vortex occurs in the solution. The solution is covered
and allowed to sit for 7 hours at room temperature. Solution A is
then added to a mixing vessel and mixed with 50.70 wt. % of NeoCryl
A 2092 and 1.0 wt. % defoamer (BYK-012) with a Cowels dispersion
blade until smooth and lump free. Added under agitation is 8.91 wt.
% of carbazole violet 8406E and 2.49 wt. % of White Mica 6300 while
blending using a Cowels disperser. The solution is mixed for 15
minutes and the viscosity is checked. To achieve the desired
viscosity, additional methyl ethyl hydroxyethyl cellulose
(available as Bermocoll 5500 from AkzoNobel) may be added to the
mixture under vigorous agitation. The viscosity is 105 Krebs units
and the solution is allowed to cool to room temperature. Violet ink
prepared as illustrated above was measured on an Agilant 8451
UV-Vis spectrophotometer and diluted until the peak transmission
shown in FIGS. 5 and 6 was produced. The violet ink composition had
a peak transmission at 451 nm.
Example C
A violet ink composition is prepared. Solution A (Thickener) is
prepared using 37.60 wt. % of deionized water and 1.0 wt. % of
ethyl hydroxyethyl cellulose (Bermocoll EBM 5500--Akzo Nobel). The
deionized water is added to a mixing vessel and the cellulose is
gradually added to the water under agitation. A Cowels dispersion
blade is used for mixing. The solution is mixed for 25 minutes
until smooth and lump free. AMP 95 in an amount of 0.05 wt. % is
then added and mixed into solution. After initial thickening of the
solution, it is further mixed for 30 minutes, increasing the mixing
speed as the thickening occurs. The speed of mixing is adjusted so
that a vortex occurs in the solution. The solution is covered and
allowed to sit overnight at room temperature. Solution A is then
added to a mixing vessel and mixed with 49.51 wt. % of NeoCryl A
2092 and 0.9 wt. % BYK-012 with a Cowels dispersion blade until
smooth and lump free. Added under agitation is 7.91 wt. % of
carbazole violet 8406E and 3.49 wt. % of white mica 6300 while
blending using a Cowels disperser. The solution is mixed for 20
minutes and the viscosity is checked. To achieve the desired
viscosity, additional methyl ethyl hydroxyethyl cellulose
(available as Bermocoll 5500 from AkzoNobel) may be added to the
mixture under vigorous agitation. The viscosity is 95 Krebs units
and the solution is allowed to cool to room temperature. Violet ink
prepared as illustrated in this example was on an Agilant 8451
UV-Vis spectrophotometer and diluted until a peak transmission was
obtained, shown in FIGS. 5 and 6. The violet ink composition had a
peak transmission at 451 nm.
Example D
A green ink is prepared using 25.61 wt. % of deionized water and
0.8 wt. % of ethyl hydroxyethyl cellulose (Bermocoll EBM 5500--Akzo
Nobel). The deionized water is added to a mixing vessel and the
cellulose is gradually added to the water under agitation. A Cowels
dispersion blade is used for mixing. The solution is mixed for 25
minutes until smooth and lump free. AMP 95 in an amount of 0.05 wt.
% is then added and mixed into solution. After initial thickening
of the solution, it is mixed for 30 minutes, increasing the mixing
speed as it thickens until a vortex appeared in the solution. The
solution is covered and allowed to sit at room temperature for five
hours and is then added to a mixing vessel and mixed with 35.33 wt.
% of NeoCryl A 2092 and 0.73 wt. % BYK-011 defoamer and blended
with a Cowels dispersion blade until smooth and lump free. Pigments
in the amounts of 26.98 wt. % of Phtahalo Green YS 5166E and 10.1
wt. % of white mica 6300 and 0.4 wt. % Euxyl PE-9010 were added
under agitation while blending using a Cowels disperser. The
solution is mixed for 20 minutes and the viscosity is checked. To
achieve the desired viscosity, additional methyl ethyl hydroxyethyl
cellulose (available as Bermocoll 5500 from AkzoNobel) may be added
to the mixture under vigorous agitation. The viscosity is 95 Krebs
units and the solution is allowed to cool to room temperature.
Green ink prepared as illustrated above was measured on an Agilant
8451 UV-Vis spectrophotometer and diluted until a peak transmission
was, as shown in FIGS. 5 and 6, was obtained. The green ink
composition had a peak transmission at 545 nm.
Example E
A green ink is prepared using 27.06 wt. % of deionized water and
0.7 wt. % of ethyl hydroxyethyl cellulose (Bermocoll EBM 5500--Akzo
Nobel). The deionized water is added to a mixing vessel and the
cellulose is gradually added to the water under agitation. A Cowels
dispersion blade is used for mixing. The solution is mixed for 30
minutes until smooth and lump free. AMP 95 in an amount of 0.05 wt.
% is then added and mixed into solution. After initial thickening
of the solution, it is mixed for 25 minutes, increasing the mixing
speed as it thickens until a vortex appeared in the solution. The
solution is covered and allowed to sit at room temperature for
eight hours and is then added to a mixing vessel and mixed with
34.83 wt. % of NeoCryl A 2092 and 0.9 wt. % BYK-011 defoamer and
blended with a Cowels dispersion blade until smooth and lump free.
Added under agitation while blending with a Cowels disperser are
26.48 wt. % of Phtahalo Green YS 5166E and 9.6 wt. % of white mica
6300 and 0.5 wt. % Euxyl PE-9010. The solution is mixed for 20
minutes and the viscosity is checked. To achieve the desired
viscosity, additional methyl ethyl hydroxyethyl cellulose
(available as Bermocoll 5500 from AkzoNobel) may be added to the
mixture under vigorous agitation. The viscosity is 100 Krebs units
and the solution is allowed to cool to room temperature. Green ink
prepared as illustrated above was measured on an Agilant 8451
UV-Vis spectrophotometer and diluted until a peak transmission was,
as shown in FIGS. 5 and 6, was obtained. The green ink composition
had a peak transmission at 545 nm.
Example F
A blue ink composition is prepared using 29.57 wt. % of deionized
water and 0.75 wt. % of ethyl hydroxyethyl cellulose (Bermocoll EBM
5500--Akzo Nobel). The deionized water is added to a mixing vessel
and the cellulose is gradually added to the water under agitation.
A Cowels dispersion blade is used for mixing. The solution is mixed
for 25 minutes until smooth and lump free. AMP 95 in an amount of
0.05 wt. % is then added and mixed into solution. After initial
thickening of the solution, it is mixed for 30 minutes, increasing
the mixing speed to compensate for the thickening of the solution
until a vortex is in the solution. The solution is covered and
allowed to sit at room temperature for five hrs. and is then added
to a mixing vessel and mixed with 37.61 wt. % of NeoCryl A 2092 and
0.83 wt. % BYK-011 Defoamer and blended with a Cowels dispersion
blade until smooth and lump free. Added under agitation while
blending with a Cowels disperser are 23.28 wt. % of Phtahalo Blue
GS 4820E and 7.53 wt. % of white mica 6300. A preservative in the
amount of 0.45 wt. % of Euxyl PE-9010 is also added. The solution
is mixed for 25 minutes and the viscosity is checked. To achieve
the desired viscosity, additional methyl ethyl hydroxyethyl
cellulose (available as Bermocoll 5500 from AkzoNobel) may be added
to the mixture under vigorous agitation. The viscosity is 103 Krebs
units and the solution is allowed to cool to room temperature. Blue
ink prepared as illustrated in this example was measured on Agilant
8451 UV-Vis spectrophotometer and diluted until the peak
transmission shown in FIGS. 5 and 6 was obtained. The blue ink
composition had a peak transmission at 506 nm.
Example G
A blue ink composition is prepared using 31.57 wt. % of deionized
water and 0.55 wt. % of ethyl hydroxyethyl cellulose (Bermocoll EBM
5500--Akzo Nobel). The deionized water is added to a mixing vessel
and the cellulose is gradually added to the water under agitation.
A Cowels dispersion blade is used for mixing. The solution is mixed
for 35 minutes until smooth and lump free. AMP 95 in an amount of
0.1 wt. % is then added and mixed into solution. After initial
thickening of the solution, it is mixed for 30 minutes, increasing
the mixing speed to compensate for the thickening of the solution
until a vortex is in the solution. The solution is covered and
allowed to sit at room temperature for seven hours and is then
added to a mixing vessel and mixed with 38.94 wt. % of NeoCryl A
2092 and 0.68 wt. % BYK-011 Defoamer and blended with a Cowels
dispersion blade until smooth and lump free. Added under agitation
while blending with a Cowels disperser are 21.28 wt. % of Phtahalo
Blue GS 4820E and 6.53 wt. % of white mica 6300. A preservative in
the amount of 0.35 wt. % of Euxyl PE-9010 is also added. The
solution is mixed for 25 minutes and the viscosity is checked. To
achieve the desired viscosity, additional methyl ethyl hydroxyethyl
cellulose (available as Bermocoll 5500 from AkzoNobel) may be added
to the mixture under vigorous agitation. The viscosity is 110 Krebs
units and the solution is allowed to cool to room temperature. Blue
ink prepared as illustrated in this example was measured on Agilant
8451 UV-Vis spectrophotometer and diluted until the peak
transmission shown in FIGS. 5 and 6 was obtained. The blue ink
composition had a peak transmission at 506 nm.
Example H
A red ink composition is prepared using 32.70 wt. % of deionized
water and 0.83 wt. % of ethyl hydroxyethyl cellulose (Bermocoll EBM
5500--Akzo Nobel). The deionized water is added to a mixing vessel
and the cellulose is gradually added to the water under agitation.
A Cowels dispersion blade is used for mixing. The solution is mixed
for 25 minutes until smooth and lump free. 0.05 wt. % AMP 95 is
then added and mixed into solution. After initial thickening of the
solution, it is mixed for 25 minutes, increasing the mixing speed
to compensate for the thickening of the solution until a vortex is
in the solution. The solution is covered and allowed to sit at room
temperature for twelve hours and is then added to a mixing vessel
and mixed with 41.6 wt. % of NeoCryl A 2092 and 0.91 wt. % BYK-012
defoamer and blended with a Cowels dispersion blade until smooth
and lump free. Added under agitation while blending with a Cowels
disperser are 17.18 wt. % of Naphthol Red 7949E and 6.4 wt. %
Quinacridone Red 8030E. A preservative in the amount of 0.45 wt. %
of Euxyl PE-9010 is also added. The solution is mixed for 30
minutes and the viscosity is checked. To achieve the desired
viscosity, additional methyl ethyl hydroxyethyl cellulose
(available as Bermocoll 5500 from AkzoNobel) may be added to the
mixture under vigorous agitation. The viscosity is 110 Krebs units
and the solution is allowed to cool to room temperature. Red ink
prepared as illustrated in this example was measured on an Agilant
8451 UV-Vis spectrophotometer and diluted until the peak
transmission shown in FIGS. 5 and 6 was obtained. The red ink
composition had two peak transmissions at 449 nm and 716 nm.
Example I
A red ink composition is prepared using 34.70 wt. % of deionized
water and 1.03 wt. % of ethyl hydroxyethyl cellulose (Bermocoll EBM
5500--Akzo Nobel). The deionized water is added to a mixing vessel
and the cellulose is gradually added to the water under agitation.
A Cowels dispersion blade is used for mixing. The solution is mixed
for 25 minutes until smooth and lump free. 0.05 wt. % AMP 95 is
then added and mixed into solution. After initial thickening of the
solution, it is mixed for 25 minutes, increasing the mixing speed
to compensate for the thickening of the solution until a vortex is
in the solution. The solution is covered and allowed to sit at room
temperature for six hours and is then added to a mixing vessel and
mixed with 43.43 wt. % of NeoCryl A 2092 and 0.76 wt. % BYK-012
defoamer and blended with a Cowels dispersion blade until smooth
and lump free. Added under agitation while blending with a Cowels
disperser are 15.18 wt. % of Naphthol Red 7949E and 4.4 wt. %
Quinacridone Red 8030E. A preservative in the amount of 0.45 wt. %
of Euxyl PE-9010 is also added. The solution is mixed for 30
minutes and the viscosity is checked. To achieve the desired
viscosity, additional methyl ethyl hydroxyethyl cellulose
(available as Bermocoll 5500 from AkzoNobel) may be added to the
mixture under vigorous agitation. The viscosity is 105 Krebs units
and the solution is allowed to cool to room temperature. Red ink
prepared as illustrated in this example was measured on an Agilant
8451 UV-Vis spectrophotometer and diluted until the peak
transmission shown in FIGS. 5 and 6 was obtained. The red ink
composition had two peak transmissions at 449 nm and 716 nm.
Example J
An orange ink composition is prepared using 35.96 wt. % of
deionized water and 0.96 wt. % of ethyl hydroxyethyl cellulose
(Bermocoll EBM 5500--Akzo Nobel). The deionized water is added to a
mixing vessel and the cellulose is gradually added to the water
under agitation. A Cowels dispersion blade is used for mixing. The
solution is mixed for 30 minutes until smooth and lump free and
0.05 wt. % AMP 95 is then added and mixed into solution. After
initial thickening of the solution, it is mixed for an additional
25 minutes, increasing the mixing speed to compensate for the
thickening of the solution until a vortex is in the solution. The
solution is covered and allowed to sit at room temperature for
twelve hours and is then added to a mixing vessel and mixed with
46.64 wt. % of NeoCryl A 2092 and 1.05 wt. % BYK-011 defoamer and
blended with a Cowels dispersion blade until smooth and lump free.
Added under agitation while blending with a Cowels disperser are
6.22 wt. % of Benzimidazolone Yellow 5696E, 6.16 wt. %
Disazopyrazolone Orange 6012E and 2.5 wt. % White Mica 6300. Euxyl
PE-9010 preservative in the amount of 0.53 wt. % is also added. The
solution is mixed for 15 minutes and the viscosity is checked. To
achieve the desired viscosity, additional methyl ethyl hydroxyethyl
cellulose (available as Bermocoll 5500 from AkzoNobel) may be added
to the mixture under vigorous agitation. The viscosity is 90 Krebs
units and the solution is allowed to cool to room temperature.
Orange ink prepared as illustrated in this example was measured on
an Agilant 8451 UV-Vis spectrophotometer and diluted until the peak
transmission shown in FIGS. 5 and 6 was obtained. The orange ink
composition had two peak transmissions at 352 nm and 636 nm.
Example K
An orange ink composition is prepared using 37.96 wt. % of
deionized water and 1.10 wt. % of ethyl hydroxyethyl cellulose
(Bermocoll EBM 5500--Akzo Nobel). The deionized water is added to a
mixing vessel and the cellulose is gradually added to the water
under agitation. A Cowels dispersion blade is used for mixing. The
solution is mixed for 30 minutes until smooth and lump free and
0.10 wt. % AMP 95 is then added and mixed into solution. After
initial thickening of the solution, it is mixed for an additional
25 minutes, increasing the mixing speed to compensate for the
thickening of the solution until a vortex is in the solution. The
solution is covered and allowed to sit at room temperature for
twelve hours and is then added to a mixing vessel and mixed with
48.64 wt. % of NeoCryl A 2092 and 1.20 wt. % BYK-011 defoamer and
blended with a Cowels dispersion blade until smooth and lump free.
Added under agitation while blending with a Cowels disperser are
4.91 wt. % of Benzimidazolone Yellow 5696E, 4.16 wt. %
Disazopyrazolone Orange 6012E and 1.5 wt. % White Mica 6300. Euxyl
PE-9010 preservative in the amount of 0.43 wt. % is also added. The
solution is mixed for 15 minutes and the viscosity is checked. To
achieve the desired viscosity, additional methyl ethyl hydroxyethyl
cellulose (available as Bermocoll 5500 from AkzoNobel) may be added
to the mixture under vigorous agitation. The viscosity is 95 Krebs
units and the solution is allowed to cool to room temperature.
Orange ink prepared as illustrated in this example was measured on
an Agilant 8451 UV-Vis spectrophotometer and diluted until the peak
transmission shown in FIGS. 5 and 6 was obtained. The orange ink
composition had two peak transmissions at 352 nm and 636 nm.
Example L
An orange ink composition is prepared using 33.05 wt. % of
deionized water and 0.88% wt. % of ethyl hydroxyethyl cellulose
(Bermocoll EBM 5500--Akzo Nobel). The deionized water is added to a
mixing vessel and the cellulose is gradually added to the water
under agitation. A Cowels dispersion blade is used for mixing. The
solution is mixed for 30 minutes until smooth and lump free and
0.10 wt. % AMP 95 is then added and mixed into solution. After
initial thickening of the solution, it is mixed for an additional
35 minutes, increasing the mixing speed to compensate for the
thickening of the solution until a vortex is in the solution. The
solution is covered and allowed to sit at room temperature
overnight and is then added to a mixing vessel and mixed with
42.95% wt. % of NeoCryl A 2092 and 0.65 wt. % BYK-011 defoamer and
blended with a Cowels dispersion blade until smooth and lump free.
Added under agitation while blending with a Cowels disperser are
11.79% wt. % of Benzimidazolone Yellow 5696E, 2.36 wt. %
Disazopyrazolone Orange 6012E and 7.72 wt. % Orange 43 6002E. Euxyl
PE-9010 preservative in the amount of 0.55 wt. % is also added. The
solution is mixed for 20 minutes and the viscosity is checked. To
achieve the desired viscosity, additional methyl ethyl hydroxyethyl
cellulose (available as Bermocoll 5500 from AkzoNobel) may be added
to the mixture under vigorous agitation. The viscosity is 90 Krebs
units and the solution is allowed to cool to room temperature.
Orange ink prepared as illustrated in this example is measured on
an Agilant 8451 UV-Vis spectrophotometer and diluted until a peak
transmission is obtained. It is predicted that the orange ink
composition will have two peak transmissions between 450 nm and 675
nm.
Example M
An orange ink composition is prepared using 35.05 wt. % of
deionized water and 0.65% wt. % of ethyl hydroxyethyl cellulose
(Bermocoll EBM 5500--Akzo Nobel). The deionized water is added to a
mixing vessel and the cellulose is gradually added to the water
under agitation. A Cowels dispersion blade is used for mixing. The
solution is mixed for 25 minutes until smooth and lump free and
0.10 wt. % AMP 95 is then added and mixed into solution. After
initial thickening of the solution, it is mixed for an additional
30 minutes, increasing the mixing speed to compensate for the
thickening of the solution until a vortex is in the solution. The
solution is covered and allowed to sit at room temperature for
seven hours and is then added to a mixing vessel and mixed with
44.95% wt. % of NeoCryl A 2092 and 0.5 wt. % BYK-011 defoamer and
blended with a Cowels dispersion blade until smooth and lump free.
Added under agitation while blending with a Cowels disperser are
12.22% wt. % of Benzimidazolone Yellow 5696E, 0.36 wt. %
Disazopyrazolone Orange 6012E and 5.72 wt. % Orange 43 6002E. Euxyl
PE-9010 preservative in the amount of 0.45 wt. % is also added. The
solution is mixed for 20 minutes and the viscosity is checked. To
achieve the desired viscosity, additional methyl ethyl hydroxyethyl
cellulose (available as Bermocoll 5500 from AkzoNobel) may be added
to the mixture under vigorous agitation. The viscosity is 100 Krebs
units and the solution is allowed to cool to room temperature.
Orange ink prepared as illustrated in this example is measured on
an Agilant 8451 UV-Vis spectrophotometer and diluted until a peak
transmission is obtained. It is predicted that the orange ink
composition will have two peak transmissions between 450 nm and 675
nm.
Example N
A yellow ink composition is prepared using 31.01 wt. % of deionized
water and 0.78 wt. % of ethyl hydroxyethyl cellulose (Bermocoll EBM
5500--Akzo Nobel). The deionized water is added to a mixing vessel
and the cellulose is gradually added to the water under agitation.
A Cowels dispersion blade is used for mixing. The solution is mixed
for 20 minutes until smooth and lump free and 0.05 wt. % AMP 95 is
then added and mixed into solution. After initial thickening of the
solution, it is mixed for an additional 25 minutes, increasing the
mixing speed as the solution thickens until a vortex is appears.
The solution is covered and allowed to sit at room temperature for
twelve hours and is then added to a mixing vessel and mixed with
39.45 wt. % of NeoCryl A 2092 and 0.96 wt. % BYK-012 defoamer and
blended with a Cowels dispersion blade until smooth and lump free.
Added under agitation while blending with a Cowels disperser is
27.63 wt. % of Benzimidazolone Yellow 5696E. Euxyl PE-9010
preservative in the amount of 0.49 wt. % is also added. The
solution is mixed for 20 minutes and the viscosity is checked. To
achieve the desired viscosity, additional methyl ethyl hydroxyethyl
cellulose (available as Bermocoll 5500 from AkzoNobel) may be added
to the mixture under vigorous agitation. The viscosity is 95 Krebs
units and the solution is allowed to cool to room temperature.
Yellow ink prepared as illustrated in this example was measured on
an Agilant 8451 UV-Vis spectrophotometer and diluted until the peak
transmission shown in FIGS. 5 and 6 was obtained. The yellow ink
composition had two peak transmissions at 322 nm and 605 nm.
Example O
A yellow ink composition is prepared using 33.0 wt. % of deionized
water and 0.58 wt. % of ethyl hydroxyethyl cellulose (Bermocoll EBM
5500--Akzo Nobel). The deionized water is added to a mixing vessel
and the cellulose is gradually added to the water under agitation.
A Cowels dispersion blade is used for mixing. The solution is mixed
for 20 minutes until smooth and lump free and 0.05 wt. % AMP 95 is
then added and mixed into solution. After initial thickening of the
solution, it is mixed for an additional 25 minutes, increasing the
mixing speed as the solution thickens until a vortex is appears.
The solution is covered and allowed to sit at room temperature for
twelve hours and is then added to a mixing vessel and mixed with
39.14 wt. % of NeoCryl A 2092 and 1.1 wt. % BYK-012 defoamer and
blended with a Cowels dispersion blade until smooth and lump free.
Added under agitation while blending with a Cowels disperser is
25.63 wt. % of Benzimidazolone Yellow 5696E. Euxyl PE-9010
preservative in the amount of 0.49 wt. % is also added. The
solution is mixed for 20 minutes and the viscosity is checked. To
achieve the desired viscosity, additional methyl ethyl hydroxyethyl
cellulose (available as Bermocoll 5500 from AkzoNobel) may be added
to the mixture under vigorous agitation. The viscosity is 115 Krebs
units and the solution is allowed to cool to room temperature.
Yellow ink prepared as illustrated in this example was measured on
an Agilant 8451 UV-Vis spectrophotometer and diluted until the peak
transmission shown in FIGS. 5 and 6 was obtained. The yellow ink
composition had two peak transmissions at 322 nm and 605 nm.
Example P
A black ink composition is prepared using 34.53 wt. % of deionized
water and 0.8 wt. % of ethyl hydroxyethyl cellulose (Bermocoll EBM
5500--Akzo Nobel). The deionized water is added to a mixing vessel
and the cellulose is gradually added to the water under agitation.
A Cowels dispersion blade is used for mixing. The solution is mixed
for 20 minutes until smooth and lump free and 0.02 wt. % AMP 95 is
then added and mixed into solution. After initial thickening of the
solution, it is mixed for an additional 25 minutes, increasing the
mixing speed to compensate for the thickening of the solution until
a vortex is in the solution. The solution is covered and allowed to
sit at room temperature for ten hours and is then added to a mixing
vessel and mixed with 44.38 wt. % of NeoCryl A 2092 acrylic resin
and 0.90 wt. % BYK-012 defoamer and blended with a Cowels
dispersion blade until smooth and lump free. Colorant in the amount
of 17.85 wt. % of Lampblack 1625E is added under agitation while
blending using a Cowels disperser. Euxyl PE-9010 preservative in
the amount of 0.49 wt. % is also added. The solution is mixed for
15 minutes and the viscosity is checked. To achieve the desired
viscosity, additional methyl ethyl hydroxyethyl cellulose
(available as Bermocoll 5500 from AkzoNobel) may be added to the
mixture under vigorous agitation. The viscosity is 108 Krebs units
and the solution is allowed to cool to room temperature. As can be
seen in FIG. 6B, the black ink prepared as illustrated in this
example was measured on an Agilant 8451 UV-Vis spectrophotometer
and produced a lineal UV-Vis spectrum having no visible peak
transmission between 250 nm and 950 nm.
Reflective Light
All inks prepared as illustrated in Examples A through 0 were
measured for reflective light in the L.a.b. color space on an
X-Rite spectrophotometer. The L.a.b. color space is a
color-opponent space with dimension L for lightness and a and b for
the color-opponent dimensions, based on nonlinearly compressed
(e.g. CIE XYZ color space) coordinates. The scale for dimensions
"a" and "b" on the X-Rite spectrophotometer used was -100 to +100.
In dimension "a" the higher the value, the redder the color and the
lower the value the greener the color. In dimension "b" the higher
the value the more yellow the color and the lower the value the
more blue the color. Three separate measurements were taken on the
inks prepared as illustrated in Examples A through P and the values
averaged. The average values appear in Table XI below.
TABLE-US-00012 TABLE XI Average L.a.b. Color L a b Red 40.80 +49.29
+28.89 Orange 57.24 +52.04 +44.62 Orange B 53.50 +52.81 +44.52
Yellow 84.51 +6.63 +88.09 Green 53.29 -42.32 +18.8 Blue 40.94
-15.37 -30.73 Violet 31.40 +13.2 -22.13 Black 25.81 -0.02 -0.49
Sterilization Procedure
All inks in accordance with the invention may desirably be
sterilized. If sterilized via gamma radiation the inks in
accordance with the invention maintain their key characteristics of
adhering well to a wide variety of tissue types, having sufficient
viscosity such that they do not run, drip, bleed or smear onto
adjacent tissue margins or into the interior of the specimen when
the tissue is cut; adhering to tissue that has been previously
fixed in formalin before ink is applied; adhering to tissue when
the specimen is placed into formalin after inks are applied; having
colors that are recognizable and distinct from other colors under
both reflective light and transmitted light; not being detectable
on an X-ray and not leaving artifacts that are visible on imaging,
and drying when applied to tissue within 2-3 minutes or less.
TABLE-US-00013 TABLE XII Preferred Acceptable Target Range Range 33
kGy 31.0 kGy-39 kGy 27.5 kGy-45 kGy
If the inks are subjected to gamma radiation of less than 27.5 kGy
the ink may not be sterile. If the inks are subjected to gamma
radiation more than 45 kGy the ink properties may be compromised,
resulting in performance issues.
Gamma radiation comprises high-energy photons that are emitted from
an isotope source (Cobalt 60) producing ionization (electron
disruptions) throughout a product. Each lot of ink is sterilized
within approximately one week of manufacturing. When processing
each pallet is broken down and loaded into a tote of processing per
the approved load diagram and a detailed run record is followed
indicating what product, lot number and number of cases is to be
loaded in each tote. The inks in accordance with the invention are
subjected to a minimum specified dose of 27 kGy and a maximum
specified dose of 45 kGy. The target dose, and most preferred dose
is 33 kGy.
Example Q
Three inks chosen at random from ink prepared as illustrated in
Examples A through P above were applied to six areas of a piece of
tissue. The inks were applied to anterior, inferior, lateral,
medial, posterior and superior surfaces. The inks dried within two
to three minutes of application. When the specimen was X-rayed, the
ink did not leave any artifacts that were visible on the image as
best seen in FIG. 10. There was minimal to no leaching of the ink
into the formalin when tissue was submerged in the formalin. Inks
adhered securely and dried quickly without running or dripping into
tissue crevices, remaining on the surface of the tissue. In
addition, the inks did not migrate onto adjacent tissue margins.
Ink colors were bright, easily identifiable, and easily
distinguishable from one another.
Tissue is marked with the inks in accordance with the invention may
be processed with methods known to those of skill in the art. An
exemplary method of processing tissue for visualization under a
microscope is shown in Table XI.
TABLE-US-00014 TABLE XIII 10% Formalin 24-48 Hours 70% ETOH 1
Wash/30 Minutes 90% ETOH 2 Washes/30 Minutes Each 100% ETOH 2
Washes/30 Minutes Each Xylene 3 Immersions/60 Minutes Each Paraffin
2 Baths/60 Minutes Each
The tissue specimens are cut on a microtome and stained with Gills
hematoxylin and Eosin Y and are viewed under a microscope at
2.5.times. and 60.times. magnification. The inks are easily viewed
under a microscope. The inks form an opaque film or shell on the
tissue specimen outer surface with greater adherence to the tissue
than commercially available inks. The ink colors are bright, easily
identifiable, and easily distinguishable from one another.
The fixing solution composition according to the present invention
includes 95% ethanol, lactic acid, deionized water and formalin.
Preferred ranges of the foregoing are set forth in the table
below.
TABLE-US-00015 TABLE XIV FIXING SOLUTION FORMULATION Ingredient
Target Formulation Preferred Range Ethanol 95% 33.33% 31.0-35.0%
Lactic Acid 11.11% 10.1-12.1% Deionized Water 33.33% 31.3-35.3%
Formalin 22.22% 21.0-23.0%
Preparation of Fixing Solution
Example 1
Fixing solution is sometimes sprayed, dabbed, dripped, or otherwise
applied onto the inked tissue to enhance the adherence of the ink
to the tissue. The fixing solution is prepared by first mixing 31.5
wt. % of water with 34.5% wt. % ethanol (95%) in a clean stainless
steel vessel. Lactic acid in an amount of 11.5 wt. % is added
followed by 22.5 wt. % formalin while mildly stirring the mixture
for twenty to thirty minutes after which is it ready to use. The
fixing solution in accordance with the invention enhances the
adherence of the ink to the tissue as best seen in FIG. XX In
addition, the fixing solution prepared as illustrated in this
Example does not have an odor that was offensive or pungent.
Example 2
A second batch of fixative is prepared by first mixing 33.33 wt. %
of deionized water with 33.33% wt. % ethanol (95%) in a clean glass
lined vessel. Lactic acid in an amount of 11.11 wt. % is added
followed by 22.22 wt. % formalin while mildly stirring the mixture
for twenty to thirty minutes after which is it ready to use. The
fixative in accordance with the invention enhances the adhesion of
the inks to the tissue. In addition, fixative prepared as
illustrated in this Example does not have an odor that is offensive
or pungent.
Inks prepared in accordance with the invention need no further
dilution and are ready to be applied directly to tissue specimens.
When three inks are placed on a tissue specimen the inks do not
bleed onto adjacent margins and are readily visible and
distinguishable from each other under a microscope.
Although the present invention has been described with reference to
certain aspects and embodiments, those of ordinary skill in the art
will appreciate that changes may be made in form and detail without
departing from the spirit and scope of the invention.
* * * * *
References